Enhanced isolation of Serpulina hyodysenteriae by using sliced agar media

Abstract

A method has been developed for separating Serpulina hyodysenteriae, a large spirochete and the causative agent of swine dysentery (SD), from other fecal anaerobic bacteria in rectal and colonic swabs. This was done by cutting the blood agar in parallel cuts and streaking perpendicular to the cuts in the center of the petri dish. Migration of S. hyodysenteriae from the central streak was apparent by the presence of strong beta-hemolysis along the edges of the cuts. If only S. hyodysenteriae migrated in the cut, they migrated to the end of the cut. However, if both motile bacteria and S. hyodysenteriae migrated in the cut, the motile bacteria migrated to the end of the cut where they formed colonies and the S. hyodysenteriae located along the edges of the cut between the colonies of motile bacteria and the central streak. Although motile bacteria were present where S. hyodysenteriae located, the growth of the motile bacteria was partially inhibited since they rarely formed visible colonies and were low in number. The cut in the agar was thought to improve traction for the serpentine movement of the S. hyodysenteriae and for the flagellar movement of the motile bacteria. Use of sliced blond agar was superior to conventionally streaked blond agar in that (i) it was easier to see strong beta-hemolysis on sliced agar; (ii) frequently, a confirmatory diagnosis could be made using only one petri dish with sliced agar, thereby saving time and media; (iii) S. hyodysenteriae could sometimes be isolated free of other bacteria; and (iv) sliced agar was more effective in isolating S. hyodysenteriae from swine with chronic diarrhea and nondiarrhetic carriers of SD in which the shedding of S. hyodysenteriae was low.